In Vitro Thermal and Ethanol Adaptations to Improve Vinegar Fermentation at High Temperature of Komagataeibacter oboediens MSKU 3

  • Pornchanok Taweecheep
  • Kallayanee Naloka
  • Minenosuke Matsutani
  • Toshiharu Yakushi
  • Kazunobu Matsushita
  • Gunjana TheeragoolEmail author


High temperature and high ethanol concentrations obviously affect vinegar fermentation. The thermotolerant and ethanol-resistant strains are expected to become one of the technologies for effective vinegar fermentation. This study aimed to further improve thermotolerant Komagataeibacter oboediens MSKU 3 through thermal and ethanol adaptations for acetic acid fermentation. The MSKU 3 strain was independently cultured by a repetitive cultivation in gradually increasing temperature from 37 to 39 °C for thermal adaptation, while adaptation to ethanol was carried out from concentrations of 3 to 5.5% (v/v) at 37 °C. Acetic acid fermentation revealed that the thermo-adapted T4 strain could produce 2.82% acidity with 3% ethanol at 39 °C, whereas the ethanol-adapted E3 strain could produce 3.54% acidity with 5.5% ethanol at 37 °C, in contrast to the parental strain, MSKU 3, in which no fermentation occurs at either 39 °C or 5.5% ethanol. Furthermore, genome mapping analysis of T4 and E3 strains against the genome of parental strain MSKU 3 revealed several mutated genes that are associated with thermotolerance or ethanol adaptation. The occurrence of these adaptation-associated mutations during adaptive evolution was also analyzed. Therefore, adapted strains T4 and E3 revealed the potential of Komagataeibacter oboediens strain improvement to further enhance vinegar fermentation with high ethanol concentration at high temperature.


Komagataeibacter oboediens Vinegar fermentation In vitro Thermal adaptation Ethanol adaptation Mutated gene Next-generation sequencing (NGS) technology 



This work was financially supported by the Golden Jubilee PhD Program (grant no. PHD/ 0242/ 2553), Department of Microbiology and the Graduate School, Kasetsart University. A part of this work was carried out through collaboration with the Core to Core Program supported by the Japan Society for the Promotion of Science (JSPS), the National Research Council of Thailand (NRCT), Vietnam Ministry of Science and Technology, National University of Laos, University of Brawijaya, Beuth University of Applied Sciences, and The University of Manchester, and also supported financially by the Advanced Low Carbon Technology Research and Development Program (ALCA: JPMJAL1106) of Japan Science and Technology Agency (JST). We would like to express our sincere thanks to Assistant Professor Dr. Naoya Kataoka, Yamaguchi University, for useful suggestions.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

12010_2019_3003_MOESM1_ESM.docx (257 kb)
Fig S1 Time course of growth and acetic acid production of K. oboediens MSKU 3 (●) and K. europaeus DSM 6160T (○) grown in 100 ml of YPGD1A2E by fed batch fermentation. The cultures were incubated at 30 °C with a rotary shaking speed of 200 rev min−1 for 14 days. The arrows indicate the new fermentation cycle started by adding 2% (v/v) ethanol, when the ethanol content is between 0.5–1.0%. Black and white arrows indicate the ethanol adding points for K. oboediens MSKU 3 and K. europaeus DSM 6160T, respectively. (DOCX 256 kb)
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ESM 1 (DOCX 13 kb)
12010_2019_3003_MOESM3_ESM.docx (19 kb)
ESM 2 (DOCX 18 kb)
12010_2019_3003_MOESM4_ESM.docx (21 kb)
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Authors and Affiliations

  1. 1.Interdisciplinary Graduate Program in Genetic Engineering, The Graduate SchoolKasetsart UniversityBangkokThailand
  2. 2.Department of Microbiology, Faculty of ScienceKasetsart UniversityBangkokThailand
  3. 3.Graduate School of Science and Technology for InnovationYamaguchi UniversityYamaguchiJapan
  4. 4.Department of Biological Chemistry, Faculty of AgricultureYamaguchi UniversityYamaguchiJapan
  5. 5.Research Center for Thermotolerant Microbial ResourcesYamaguchi UniversityYamaguchiJapan

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